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Influence of material compliance on human corneal stromal cell behaviour
Author(s) -
Kelly C.,
Ahearne M.
Publication year - 2016
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2016.0336
Subject(s) - stromal cell , materials science , dapi , polydimethylsiloxane , focal adhesion , adhesion , biomedical engineering , cell adhesion , substrate (aquarium) , biophysics , cornea , staining , cell , chemistry , nanotechnology , composite material , pathology , ophthalmology , biology , medicine , ecology , biochemistry
Purpose Due to a shortage of healthy donor tissue, bioengineering strategies for corneal regeneration are drawing more attention. It is well established that substrate stiffness has an effect on the proliferation and phenotype of many cell types. In this study, we aim to investigate the influence of the mechanical environment on corneal cell behaviour, by demonstrating the effects of varying material stiffness on the growth and morphology of human corneal stromal cells. Methods Using blends of two commercially available polydimethylsiloxane (PDMS) elastomer kits, cured polymer substrates with elastic moduli ranging from 12 kPa to 3 MPa were achieved. Materials underwent tensile mechanical testing to 10% strain in order to determine elastic mechanical properties. Samples were coated with a thin layer of collagen I to promote cell adhesion. Results Contact angle measurements were taken before and after protein coating, with a decrease in angle from 90.30 ± 0.47° to 73.26 ± 2.24°, indicating a significant increase in hydrophilicity of the PDMS. Phase contrast imaging was used to visualise differences in morphology of cells seeded onto the substrates, while actin and DAPI (4′,6‐diamidino‐2‐phenylindole) staining were employed to view differences in cytoskeletal structure under confocal microscopy. Cells on softer substrates displayed increased proliferation, as determined by AlamarBlue assay, with a notably spread and elongated fibroblastic morphology. Stiffer substrates induced increased cell aggregates, with a more dendritic shape visible. Gene expression of cells was also monitored by qPCR, with observed upregulation of collagen I, ALDH3A1 and keratocan, which was more pronounced in cells seeded on less compliant substrates. Conclusions These results will be useful in future biomaterial scaffold designs for corneal tissue regeneration.